Method of and pledget for the treatment of accessible hemorrhage with snake venom having blood coagulating properties in vitro



Patented June 14, 1938 PATENT OFFICE 2,120,680 METHOD or AND PLEDGET FOR. THE

TREATMENT OF ACCESSIBLE HEMOR- RHAGE WITH SNAKE VENOM HAVING BLOOD VITRO CQAGULATING PROPERTIES Samuel Rosenfeld and Sidney Edward Lenke, Brooklyn, N. Y., assignors to Mark A. Lenke, Port Washington, N. Y.

No Drawing. Application July 13, 1935,

Serial No. 31,178

10 Claims. (ol. 167-74) The present invention relates to a method of and article for the treatment of bleeding body tissues to arrest and stop the flow of blood therefrom, and it has particular relation to a new and improved method of and article for the treatment of such bleeding body tissues with snake venoms known to have blood coagulating properties and characteristics in vitro.

It is well known that certain types of poisonous snake venoms have properties and characteristics capable of producing the coagulation of blood in,

vitro. The more important snakes, whose venoms are recognized as having blood coagulating properties in vitro, are the tiger snake (Notechzs vscutatus) and the black snake (Pseudechz's i orphg riaceus) of the Colubridaeiamily; and the phoorsa (Echis carz'natus), urutu (Lachesis neuwiedii), fer-de-lance (Lachesis lanceolatus or bothrops a trox) and the Daboia (Vipera russellii) oi. Viperidae family. The venoms of these particular snakes have been diluted in a saline solution and tested in vitro for their blood coagulating properties and characteristics and in 'vivo in' laboratory animals, but never in the human body. 'The action by which these venoms stop hemorrhage is by producing a rapid clotting of the blood flowing through the bleeding tissues.

Heretofore, solutions of these snake venoms have been used for the preparation of antivenoms, which in turn are used to counteract a snake bite. In most' of these cases the antivenom is made from thesame venom for which it is intended to be employed as an antidote.

Certain other snake venoms, such the copperhead and the cotton mouth moccasin have been employed by subcutaneous and intravenous injection at sites distant from the bleeding tissues to prevent or cure bleeding diathesis. These particular venoms have no blood coagulating action in vitro, and their modus operandi is quite different and distinct from the venoms having blood coagulatingproperties in vitro. The action of the copperhead and cotton mouth moccasin is believed due to the fact that these venoms raises the resistance of the blood vessels to hemorrhage by an eflect directly upon the blood vessels or by the production of circulating bodies (antihemorrhagin) or a combination of the two efl'ects.

The present invention is the result of repeated eflorts to utilize the blood coagulating properties of the snake venoms which test favorably in vitro without subjecting the patient to the imdesirable neurotoxin thereof, and to provide a safe, efficient and eflective method of and means for utilizing such poisonous venoms in the treatment of bleeding tissues without subjecting the patient to such neurotoxin dangers.

One of the objects of the present invention is to provide a new and improved method of treatment of bleeding body tissues with snake venoms to quickly and effectively arrest and stop the flow of blood therefrom.

Another object of the invention is the provision of a blood coagulant of the venomous type in the form of a colorless and odorless liquid, free from bacteria formation and growth, which can be readily and quickly applied to the bleeding tissue to arrest and stop the flow of blood therefrom.

A further object of the invention is the provision of a blood coagulant of the venomous type in the form of an odorless powder, which can be quickly dissolved into a solution to be used to arrest and stop the flow of blood from bleeding body tissues with a minimum of time and trouble.

A further object-ofthe invention is to provide a pledget impregnated with a snake venom soluonly the addition of distilled water prior to its application to the bleeding tissue to produce a.

pledget capable of causing the arrest and stop page of the flow of blood therefrom.

In carrying out the invention, the fluid snake venom is first dried and then pulverized in any conventional manner. A sterile normal saline solution is then prepared, which consists of distilled water to which is added approximately nine-tenths percent sodium chloride by weight and stirred until entirely dissolved. The dried snake venom is then dissolved in the normal saline solution in the ratio of one gram of dried v snake venom to not less than one thousand cubic centimeters of the saline solution. In actual practice, however, it has been found that these proportions may be varied somewhat without detracting from the benefits to be derived therefrom. Dilutions of one gram of snake venom to one hundred cubic centimeters of diluent, and one gram of snake venom to one hundred thousand cubic centimeters of diluent have been used with success. However, if the solutions are made too.

strong, they will manifestly contain a greatif amount of neurotoxin, andjthe patient is likely to ent, and that the presence or .addition of the saline solution willnot prevent the formation and growth of these pathogens or other bacteria. In view of these spores, it has been found necessary and advisable to add to the saline solution, containing the venom, an antiseptic sufiicient to prevent the formation of and kill the growth of any bacteria therein and at the same time be innocuous to the venom. Several antiseptics, such as merthiolate, phenol, and trikresoL'have been found to be satisfactory for the purpose. In the use of merthiolate a one percent solution is added to the diluent in the ratio of one to one hundred; in the use of phenol we recommend a ratio by volume of one-tenth of one per cent of diluent; and in the use of trikresol the addition of three-tenths of onepercent of dilulent is suggested. Sterilization of the solution by either ultrafiltration or by heat seriously impairs the clotting power of the venoms, and therefore they can not be employed advantageously for the purpose. v

It has been found that the venoms deteriorate rather rapidly, insofar as their clotting power is concerned, when they are maintained for any length of time in solution form, but that the dry venom may be kept indefinitely without endangering its clotting power as a blood coagulating medium. Therefore, it has been found advisable, for commercial purposes, to prepare and maintain the venom in a dry form by means of a car'- rier powder; An innocuous powder for this purpose is lactose. A satisfactory method of preparing the fluid venom inthis manner is to dry it thoroughly, and then pulverizethe driedproduct before; admixing it with lactose in the ratio of one gram of venom to one hundred and fifty grams of lactose, After mixing the powder or admixture should be placed in a hermetically sealed or otherwise sterilecontainer. To use'the powder upon its removal from the container, it should be dissolved in anormal' saline solution in the above ratio, including an antiseptic, and

applied to the bleeding tissue by means of a pledget. v r

The most desirable method 'ofapplying .the

venomous solutions to the bleeding tissues is by means of a pledget that has been treated or soaked with the venomous solution. The pledget,

.ing ceases.

of teeth,

which may consist of a piece of gauze, folded or unfolded, cotton, silk, lint or asbestos packing, etc., is first soaked with the venomous saline solution and applied wet, topically, to the bleeding tissue, which may be any cut or open wound on the body, internal cavity caused from the removal pressed against the bleeding tissue until the bleed- Should the pledget be left pressed against the tissue for any length of time, it may have a tendency to dry is normally considered or any internal bleeding tissue made v accessible by an incision. The wet pledget is.

out in which event it i may again be soaked with the solution or removed and a new wetpledget substituted therefor. A dry pledget will have the same clotting effect, but because it is dry it will have a tendency to stick to the fibrin clot and may pull the clot free from the skin upon attempting to remove the pledget, thereby reopening the tissue and causing a renewal of the bleeding. For this reason and this reason only a wet pledget is preferable.

Since, as above explained, the venoms do not have a tendency to deteriorate when maintained in a dry condition, it has been found advisable ,and practical to prepare the pledget and distribute it commercially in a dry form. To prepare such a pledget, it is first thoroughly soaked or impregnated with the venom solution above de scribed and then allowed to dry until the moisture is removed by means of evaporation, which leaves the solids, snake venom, salt and antiseptic materials, behind in the interstices of the pledget. The pledget may now be packaged and wrapped ready for distribution and sale. Upon removal of the pledget for use, it may readily be made into a wet pledget for application to a bleeding tissue by adding distilled water in an amount necessary to thoroughly soak the same, which water is approximately equal in proportion to that removed by the evaporation process.

. Pledgets treated with snake venoms in the manner'hereinabove described have been used in the actual treatment of severe cases of hemorrhagic diathesis, dentalhemorrhage, blood dyscrasia, and hemophilia with complete success.

' In these cases the venom pledget was not resorted to. until every known means of controlling the hemorrhage was employed. After futile attempts to control the bleeding with tannic acid, adrenalin packings, thromboplastin packings and. electrocautery, a snake venom pledget was applied and the-bleeding ceased within a period of from two to three-minutes after the first application. Other cases, suffering from'intractable post-extractal hemorrhage, which had previously been treated over long periods of time by frequent injections of. the cotton mouth moccasin venom without success, were treated with avenom pledget of the character described and the fiow of blood was stoppedalmost immediately. In some of these cases, however, the bleeding reappeared again aftera period of severalhours after removal of the. pledget, or immediately upon re-' moval of the same, but in each of these instances the hemorrhage was stopped upon the application of another venom pledget. Neurological examinations of the patientsthat were treated with the snake venom pledget disclosed no evidence that the neurotoxin of the venom was being absorbed.

When we refer to snake venoms having blood coagulating properties and characteristics in vitro, we meana venom that when added to fresh formation of a clot therein.- In our experiments we determine the clotting power of any venom by the use of an arbitrary unit system. Under our system, the venom is dissolved in a saline solution at a known dilution, and a row of test tubes is set up' and by pipette means a, system is established wherein each test tube contains a dilution of venom twice as weak as the preceding tube; To each test tube is then added a fixed amount of fresh citrated blood plasma, and the time between the addition of the plasma and the formation of the clot in each tube is" measured. The final clotting unit is based upon the dilution of venom'of the greatest weakness which will citrated blood plasma in vitro will cause the cause the plasma to clot in a period of about two or three minutes, thereby keeping well within clinical requirements.

Although we have only described several modifications which our invention may assume, it will be apparent to those skilled in the art that the invention is not so limited, but that various other modifications may be made therein without departing from the spirit thereof or from the scope of the appended claims.

What we claim is:

1. A method of manufacturing a pledget to be used in the topical treatment of bleeding body tissue to cause the arrest and stoppage of the flow of blood therefrom, which comprises wetting the pledget with a solution containing snake venom having blood coagulating properties in vitro.

2. A method of manufacturing a pledget to be used in the treatment of bleeding body tissue to arrest the fiow of blood therefrom, which comprises soaking the pledget in a solution containing snake venom having blood coagulating properties in vitro, and then drying the same to remove the moisture therefrom.

3. I'he method of manufacturing a pledget to be used in the treatment of bleeding body tissue v by direct application thereto, which comprises immersing the pledget in a solution containing a snake venom having blood coagulating properties in vitro and including an antiseptic to prevent the formation and growth of bacteria in said solution, and then removing the moisture from the pledget by means of evaporation.

4. As a new article of manufacture, a pledget treated with a solution including a snake venom having blood coagulating properties and characteristics in vitro.

5. As a new article of manufacture, apledget= impregnated with a solution including a snake venom having blood coagulating characteristics in vitro diluted in a solution containing an antiseptic to prevent .the formation and growth of bacteria therein.

6. As a new article of manufacture, a pledget that has been immersed in a solution containing a snake venom having known blood coagulating properties in vitroand including an antiseptic to prevent the formation and growth of bacteria therein, and dried to remove the moisture therefrom to prevent deterioration of the blood coagulating properties of the venom.

7. The method of preparing snake venom known to have blood coagulating properties in vitro which comprises drying the fluid venom, pulverizing the dried product and then admixing the same with a dry innocuous carrier powder.

8. The method of preparing a snake venom known to have blood coagulating properties in vitro which comprises drying the fluid venom, pulverizing the dried product, and then admixing the same with a dry innocuous carrier powder containing an antiseptic to prevent the formation and growth of bacteria when the admixture is dissolved in a solution.

9. The method of preparing a snake venom known to have blood coagulating properties in vitro into a solution suitable for use in the treatment of accessible hemorrhages which comprises drying the venom, pulverizing the dried product,- and then dissolving the same in water to form a solution. H

10. The method of preparing a snake venom known to have blood coagulating properties in vitro into a solutionsuitable'for use in the treatment of accessible hemorrhages which comprises drying the venom, pulverizing the dried product, and then dissolving the same in a solution containing an antiseptic to prevent the formation and growth of bacteria therein.

SAMUEL ROSENFELD.

SIDNEY EDWARD LENKE. 

